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Cardiac commitment of primate embryonic stem cells.

Nature protocols (2008-09-06)
Julia Leschik, Sonia Stefanovic, Benjamin Brinon, Michel Pucéat
ABSTRACT

Primate nonhuman and human embryonic stem (ES) cells provide a powerful model of early cardiogenesis. Furthermore, engineering of cardiac progenitors or cardiomyocytes from ES cells offers a tool for drug screening in toxicology or to search for molecules to improve and scale up the process of cardiac differentiation using high-throughput screening technology, as well as a source of cell therapy of heart failure. Spontaneous differentiation of ES cells into cardiomyocytes is, however, limited. Herein, we describe a simple protocol to commit both rhesus and human ES cells toward a cardiac lineage and to sort out early cardiac progenitors. Primate ES cells are challenged for 4 d with the cardiogenic morphogen bone morphogenetic protein 2 (BMP2) and sorted out using anti-SSEA-1 antibody-conjugated magnetic beads. Cardiac progenitor cells can be generated and isolated in 4 d using this protocol.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Mitomycin C from Streptomyces caespitosus, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
SU5402, SU5402, CAS 215543-92-3, is a cell-permeable, reversible, and ATP-competitive inhibitor of the tyrosine kinase activity of FGFR1 (IC₅₀ = 10-20 µM in the presence of 1 mM ATP).
Sigma-Aldrich
ES Cell Characterization Kit, The Embryonic Stem Cell Characterization Kit phenotypically assesses the differentiation status of ES cells by measuring their AP activity, cell-surface stage-specific antigens (SSEA-1, SSEA-4) as well as expression of TRA-1-60, TRA-1-81 antigens.